Michael Lattke1, Robert Goldstone2, James Ellis3, Stefan Boeing4, Jeronimo Jurado-Arjona5, Nicolás Marichal5, James MacRae3, Benedikt Berninger5,6,7,8, Francois Guillemot1*

1Neural Stem Cell Biology Laboratory, The Francis Crick Institute, London, NW1 1AT, UK 2Advanced Sequencing Facility, The Francis Crick Institute, London, NW1 1AT, UK 3Metabolomics Facility, The Francis Crick Institute, London, NW1 1AT, UK 4Bionformatics & Biostatistics, The Francis Crick Institute, London, NW1 1AT, UK 5Institute of Psychiatry, Psychology & Neuroscience, Centre for Developmental Neurobiology, King's College London, London, SE1 1UL, UK 6MRC Centre for Neurodevelopmental Disorders, King's College London, London, SE1 1UL, UK
7Institute of Physiological Chemistry, University Medical Center of the Johannes Gutenberg University Mainz, 55128 Mainz, Germany 8The Francis Crick Institute, London, NW1 1AT, UK

*lead author

Corresponding author e-mail address: Francois.Guillemot@crick.ac.uk

**Contacts for this online resource:** Stefan.Boeing@crick.ac.uk, Michael.Lattke@crick.ac.uk

The underlying datasets are also available as superseries GSE152223 in the GEO database (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE152223)

Summary

Astrocytes have essential functions in brain homeostasis that are established late in differentiation, but the mechanisms underlying the functional maturation of astrocytes are not well understood. Here we identified extensive transcriptional changes that occur during murine astrocyte maturation in vivo and are accompanied by chromatin remodelling at enhancer elements. Investigating astrocyte maturation in a cell culture model revealed that in vitro-differentiated astrocytes lacked expression of many mature astrocyte-specific genes, including genes for the transcription factors Rorb, Dbx2, Lhx2 and Fezf2. Forced expression of these factors in vitro induced distinct sets of mature astrocytes-specific transcripts. Culturing astrocytes in a three-dimensional matrix containing FGF2 induced expression of Rorb, Dbx2 and Lhx2 and improved astrocyte maturity based on transcriptional and chromatin profiles. Therefore, extrinsic signals orchestrate the expression of multiple intrinsic regulators, which in turn induce in a modular manner the transcriptional and chromatin changes underlying astrocyte maturation.

Online Resource Version 0.1.4 (21-05-18)